SARS康复者M抗原特异性记忆性T细胞免疫应答的探讨

目的探讨SARS患者康复后，外周血单个核细胞（PBMCs）中是否存在SARS-CoV M抗原特异性T细胞。方法从完全康复期SAILS患者和健康人外周血中分离PBMCs，体外经M混合多肽刺激后，采用酶联免疫吸附试验（ELISA）、酶联免疫斑点试验（ELISpot）及流式细胞仪检测技术，分析抗原特异性T淋巴细胞的反应性。结果在未经任何抗原刺激的情况下，PBMCs几乎不分泌IFN-γ。当SARS-CoV M混合多肽刺激后，SARS康复期患者的PBMCs分泌大量的IFN-γ，并可检测出高频率的IFN-γ产生细胞。与健康刺激组及康复期SAILS患者未刺激组相比，差异显著（P〈0．05）。流式结果显示，SAILS康复期患者PBMCs经M多肽刺激后，分别有0．11％CD4^＋和0．16％CD8^＋T淋巴细胞分泌IFN-γ。根据IFN-γ和IL-2的表达与否，可将CD4^＋T细胞分为三个亚群：IFN-γ^-IL-2^＋、IFN-γ^＋IL-2^＋和IFN-γ^＋IL-2^-。结论SARS-CoV感染后，机体可以产生针对SARS-CoV M蛋白的抗原特异性细胞免疫反应，其免疫记忆可以在体内维持很长时间。     

汉滩病毒核衣壳蛋白C-端T细胞表位鉴定

目的　鉴定汉滩病毒核衣壳蛋白 (HTNVNP)C 端T细胞表位 ,为肾综合征出血热(HFRS)发病机理、疫苗研制及抗病毒免疫反应研究奠定基础。方法　采用Ficoll密度梯度离心法分离HFRS恢复期患者外周血单个核细胞 (PBMC)。用IFN γELISPOT实验和T细胞增殖实验 ,测试 7名患者PBMC对 2 3条NPC 端合成多肽的T细胞应答。结果　IFN γELISPOT实验结果表明 ,2名供体(3、4 )可分别检测到对 5 1、70号 2条多肽特异性T细胞应答。在供体 3,70号肽特异性T细胞频率为4 5SFC 10 6 PBMC ;在供体 4 ,5 1号肽特异性T细胞频率为 82SFC 10 6 PBMC。T细胞增殖实验与ELISPOT结果基本一致 ,但 5 3号肽和 6 4号肽还可分别刺激供体 1和供体 4的T细胞增殖 ,而未能诱导IFN γ分泌。结论　 5 1号和 70号多肽可能是NPC 端较强的T细胞表位。     

不明原因习惯性流产患者主动免疫治疗前后Th1/Th2型细胞因子水平的变化

为探讨主动免疫治疗对不明原因习惯性流产 (UHA )患者Th1/Th2型细胞因子水平的影响。采用酶联免疫吸附法检测15例正常非妊娠妇女、 35例UHA患者淋巴细胞主动免疫治疗前后经滋养细胞抗原刺激的外周血单个核细胞 (PBMC )培养上清液中IL 2、IFN γ、IL 4、IL 10的水平。结果发现 :(1)在最佳诱导时间下 ,UHA组治疗前PBMC产生IL 2、IFN γ的水平明显高于正常对照组 (P <0 0 5 ) ,IL 4、IL 10水平明显低于正常对照组 (P <0 0 5 )。UHA组治疗后PBMC产生IL 2、IFN γ的水平较治疗前明显降低 (P <0 0 5 ) ,IL 4、IL 10水平较治疗前明显升高 (P <0 0 5 )。UHA组治疗后PBMC产生各细胞因子的水平与正常对照组比较 ,差异均无显著性 (P >0 0 5 ) ;(2 )UHA组 35例患者主动免疫治疗后半年内 2 8例妊娠 ,其中 9例又出现自然流产。 9例自然流产者治疗后IL 2、IFN γ水平未明显下降 ,IL 10水平未明显上升。 19例妊娠成功者治疗后IL 2、IFN γ水平较治疗前明显下降 (P <0 0 5 ) ,IL 4、IL 10水平明显上升 (P <0 0 5 )。以上结果表明UHA患者对滋养细胞抗原产生以Th1型反应为主的免疫应答 ,产生大量Th1型细胞因子 ,主动免疫治疗有助于上调Th2型细胞因子及下调Th1型细胞因子 ,利于UHA患者妊娠成功。     

不明原因习惯性流产患者Th1/Th2型细胞因子的检测

目的 :了解不明原因习惯性流产 (UHA)患者对滋养细胞抗原刺激产生Th1、Th2型细胞因子的水平 ,探讨其在UHA发病中的作用。方法 :选取 5 4例UHA患者 ,35例为未妊娠者 ,为UHA未孕组 ,19例为有UHA病史 ,现在确定妊娠并难免流产者 ,为UHA难免流产组。分别选取 15例正常未孕妇女及正常妊娠者作为对照组。应用酶联免疫吸附法检测未孕各组经滋养细胞抗原刺激的外周血单个核细胞 (PBMC)及妊娠各组PBMC与蜕膜单个核细胞 (DMC)培养上清液中IL 2、IFN γ、IL 4、IL 10的水平。结果 :①在最佳诱导时间下 ,UHA未孕组PBMC产生IL 2、IFN γ的水平明显高于正常未孕组 (P均 <0 0 5 ) ,而IL 4、IL 10水平明显低于正常未孕组 (P均 <0 0 5 )。②UHA难免流产组PBMC及DMC产生的IL 2、IFN γ水平均明显高于正常妊娠组 (P均 <0 0 5 ) ,而IL 4、IL 10水平则明显低于正常妊娠组 ,差异均有显著性 (P均 <0 0 5 )。两组PBMC产生的各细胞因子水平均低于同组DMC ,差异有显著性 (P <0 0 5 )。结论 :UHA患者对滋养细胞抗原产生以Th1型反应为主的免疫应答 ,产生大量Th1型细胞因子 ,导致妊娠丢失。     

SARS患者康复期血清特异性抗体效价检测

目的：SARS患者康复期血清特异性抗体的效价。方法：采用间接酶联免疫法和双抗原夹心法。结果：SARS患者康复期血清特异性抗体IgG水平在住院第5周达最高值，IgM水平在住院第3周达到最高值。SARS患者康复期血清IgG水平是IgM的1～7倍。住院5～7周的SARS康复期患者，其血清特异性抗体检测阳性率达100％。结论：SAPS患者康复期血清特异性抗体水平在住院第5周达到最高值，其阳性检出率达100％。     

SARS-CoV感染者血清IFN-γ和TNF-α的动态变化及其意义

目的　探讨SARS患者血清IFN γ和TNF α在SARS冠状病毒感染免疫中的作用。方法　应用酶联免疫吸附法 (ELISA)测定广州地区 2 8例SARS冠状病毒感染患者双份血清IFN γ和TNF α的水平。实验数据采用两样本均数t检验法。结果　 2 8例SARS冠状病毒感染者急性期血清IFN γ和TNF α水平比正常健康对照组明显增高 (P <0 .0 5 ) ;恢复期血清IFN γ水平比正常健康对照组明显增高 (P <0 .0 5 )。感染病程第 1周 ,患者血清IFN γ水平达高峰 ,然后逐渐下降。SARS患者血清TNF α水平在病程第 2周达高峰 ,然后逐渐下降。结论　IFN γ和TNF α在SARS冠状病毒的致病与免疫过程中可能起重要作用     

抗原活化慢性乙型肝炎患者外周血树突状细胞的对比研究

目的 探讨不同抗原在体外活化慢性乙型肝炎患者外周血树突状细胞(DC)及诱导特异性T细胞应答的能力.方法 用无血清培养基从慢性乙型肝炎患者外周血中分离培养DC,在DC成熟前,分别加入HBsAg多肽、HBcAg多肽刺激,用流式细胞仪检测DC表型,用液闪计数仪观察DC对T细胞的增殖作用,用ELISA法检测混合淋巴细胞反应(MLR)中IL-12的分泌水平.结果 经HBcAg多肽刺激DC的CD86表达率为(92.20±5.18)%,明显高于HBsAg多肽刺激组(76.19±3.90)%和未加抗原组(62.37±4.24)%,P＜0.01;经HBcAg多肽刺激组DC诱导同种异体静止T细胞增殖的能力每分钟液闪计数值cpm为34 326±3088,明显高于HBsAg多肽刺激组20 306±2897和单个核细胞组3454±409,P＜0.01;经HBcAg多肽刺激组DC MLR中IL-12(348±42.8)ng/L,分别高于HBsAg多肽刺激组(226±30.6)ng/L和未加抗原组(116±15.6)ng/L,P＜0.01.结论 使用HBcAg多肽刺激DC可比HBsAg多肽更有效地提呈病毒抗原,提高诱导特异性T细胞应答的能力.     

SARS-CoV S DNA疫苗诱导特异性T细胞及相关细胞因子的特性研究

目的 小鼠经皮下SARS-CoV S DNA疫苗免疫后，研究其特异性T细胞及相关细胞因子的特性。方法SARS-CoV S DNA疫苗免疫BALB／c小鼠后，获取淋巴细胞悬液。经S抗原多肽刺激后，采用ELISA检测细胞培养上清液中IFN-γ／的水平，利用流式细胞仪在单个细胞水平上检测IFN-γ和IL-2的表达及其关系。结果 当S混合多肽刺激后，DNA疫苗免疫小鼠的淋巴细胞产生大量的IFN-γ，与对照鼠相比差异有统计学意义（P〈0．01）。细胞亚群分析的结果表明，IFN-γ^＋和IL-2^＋的CD4^＋T细胞百分率明显高于CD8^＋T细胞。单独产生IL-2的细胞占大多数，其次为IFN-γ和IL-2双阳性细胞，只产生IFN-γ的细胞很少。结论 SARS-CoV S DNA疫苗免疫小鼠后可以诱导抗原特异性CD4^＋和CD8^＋T细胞的产生。     

银屑病患者淋巴细胞及细胞因子在发病机制中的作用

目的 :分析银屑病患者淋巴细胞对角质形成细胞 (KC)增殖的作用 ,为有效治疗提供理论依据。方法 :分离并扩增银屑病患者皮损淋巴细胞和外周血淋巴细胞 (PBMC) ,以正常人PBMC作对照 ,与正常人KC混合培养后经MTT法检测混合培养液中的活细胞量 ,并对混合培养 48h的培养上清液进行IL 2、IL 4和IFN γ含量检测 ,经统计学处理 ,分析组间差异。结果 :银屑病患者皮损淋巴细胞及PBMC均可明显刺激KC增殖 ,其OD 5 5 0nm均值分别为 0 896± 0 110和 0 75 8± 0 0 96 ,与正常对照组比较 ,银屑病患者淋巴细胞能显著刺激KC的增殖 ,尤以皮损处淋巴细胞为甚。雷公藤多甙可抑制淋巴细胞刺激KC增殖作用。银屑病患者皮损淋巴细胞与KC混合培养细胞上清液 ,IL 2和IFN γ的浓度明显高于对照组 ,而IL 4明显低于对照组 ,IL 2和IFN γ的浓度均远高于IL 4的浓度。加入雷公藤多甙组IL 2、IFN γ和IL 4的含量与对照组无显著差异。结论 :银屑病患者淋巴细胞 (主要是Th1型细胞 )在刺激KC的增殖过程中起重要作用 ,细胞模型的建立为进一步研究银屑病的发病机制途径打下了基础。     

IL-12对慢性乙型肝炎患者PBMC产生IFN-γ、IL-10细胞因子的影响

为观察IL 12对不同临床分型慢性乙型肝炎患者外周血单个核细胞 (PBMC )产生Th1/Th2类细胞因子的影响。本实验分离 72例慢性乙型肝炎患者 (轻、中、重度 )PBMC ,分别与植物血凝素 (PHA ) (10 0 μg/ml)、HBcAg (1μg/ml)、HBeAg (1μg/ml)单独或联合IL 12 (10ng/ml)体外培养 4 8h ,ELISA法检测培养上清液中IFN γ、IL 10水平。 2 0例健康人群做对照。结果发现IL 12对PHA、HBcAg/HBeAg诱导健康人群PBMC产生Th1/Th2类细胞因子无协同效应 ,对慢性乙型肝炎患者PBMC产生IFN γ有显著协同增殖效应 ,慢性中度患者最为突出。同HBeAg联合诱导 ,不但显著增强慢性乙型肝炎患者PBMC产生IFN γ ,还抑制IL 10的产生。提示IL 12可增强慢性乙型肝炎患者IFN γ水平增高 ,但不同临床分型患者其增殖效应的强度不同 ;IL 12可促进HBeAg诱导的Th2型优势表达向Th1型优势表达转换     

Pepsinogen secretion from dispersed glands from rabbit stomach

In dispersed glands from rabbit stomach, pepsinogen secretion was stimulated by carbamylcholine, the C-terminal octapeptide of cholecystokinin (CCK-8), and structurally related peptides physalaemin and A23187 but not by bombesin or histamine. Reducing the incubation temperature from 37 degrees to 4 degrees C or adding carbonyl cyanide m-chlorophenylhydrazone reduced basal as well as stimulated pepsinogen secretion. Dibutyryl cGMP inhibited the stimulation of pepsinogen secretion caused by cholecystokinin but not that caused by carbamylcholine; atropine inhibited the stimulation of pepsinogen secretion caused by carbamylcholine but not that caused by cholecystokinin. Each cholecystokinin-related peptide had the same efficacy for stimulating pepsinogen secretion. In terms of the concentration that caused half-maximal stimulation of pepsinogen secretion, the relative potencies of cholecystokinin-related peptides were caerulein greater than CCK-8 greater than [desSO3]CCK-8 greater than gastrin. Removing extracellular calcium did not alter basal or cholecystokinin-stimulated pepsinogen secretion. These results illustrate that dispersed glands from rabbit stomach constitute a suitable preparation for examining the actions of various agents on pepsinogen secretion in vitro.     

The separation of renal activity from lung and from the venous effluent from perfused lungs

1. Antidiuretic and salt-retaining activity has been separated from cat and ox lung, and from the venous effluent of blood-perfused cat lungs, by acetone extraction, gel filtration, partition and thin-layer chromatography.2. The chromatographic properties of the renally active substance from lung and from pulmonary venous blood are similar.3. The renal actions of these substances, demonstrable in vivo and in the isolated organ, are characterized by reduction in the excretion of water and Na. Renal blood flow, glomerular filtration rate and the urinary Na/K are not significantly affected.4. Concentrations of the lung substance which equate with angiotensin II-val(5)-amide in reduction of urinary Na in rats during water diuresis and under alcohol sedation do not raise the mean arterial pressure of the anaesthetized rat nor contract the superfused rat colon. At these dose levels angiotensin II is markedly vasopressor and contracts the superfused rat colon.     

Differentiating betweenBesnoitia besnoiti from cattle andSarcocystis hoarensis from rodents

To provide a biological basis for studies designed to establish the mode of transmission of the veterinary pathogenBesnoitia besnoiti, we compared salient features of this pathogen in cattle with those ofSarcocystis hoarensis in rodents. The cysts and cystozoites of these organisms can readily be distinguished morphologically. In contrast toS. hoarensis which is well adapted to rodents,B. besnoiti fails to mature in jirds or mice and generally is lethal in jirds. Serological reagents discriminately detect these pathogens.B. besnoiti, therefore, can unambiguously be differentiated fromS. hoarensis either by morphological or serological methods or on the basis of experimental comparisons of virulence in laboratory rodents.     

谷氨酸在分离的小鼠皮层突触连接体的释放

目的 分析兴奋性神经递质谷氨酸在体外分离的小鼠皮层突触连接体的释放。 方法 解剖分离小鼠的大脑皮层,组织经匀浆后使用3层147 μm的尼龙网筛进行过滤,离心后重悬沉淀为突触连接体。谷氨酸释放检测实验根据谷氨酸脱氢酶酶促反应原理进行,利用多功能读数仪读取产物 烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的荧光值。采用谷氨酸标准品绘制标准曲线,使用制备的突触连接体进行谷氨酸释放检测,进行3次独立重复实验。结果 谷氨酸标准品浓度与荧光值具有良好的线性关系。新鲜制备的突触连接体在加入去极化剂KCl后,释放谷氨酸显著增加并逐渐达到平台期,加入Triton X-100裂解突触连接体后释放谷氨酸明显增加。冻融1次的突触连接体失去活性,与新鲜制备的突触连接体的谷氨酸释放曲线不同。 结论 对小鼠皮层突触连接体的提取方法进行了改良,检测和分析了兴奋性神经递质谷氨酸在分离制备的小鼠皮层突触连接体的释放。     

Clinicians are from Mars and pathologists are from Venus

CONTEXT: Text reports convey critical medical information from pathologists, radiologists, and subspecialty consultants. These reports must be clear and comprehensible to avoid medical errors. Pathologists have paid much attention to report completeness but have ignored the corresponding issue of report comprehension. This situation presents an increasingly serious potential problem. As laboratories are consolidated and as reports are disseminated in new ways (eg, via the World Wide Web), the target audience becomes more diverse and less likely to have any contact with pathologists beyond the written reports themselves. OBJECTIVE: To compare clinician comprehension with pathologist intent in written pathology reports. METHODS: Typical surgical pathology reports relevant to surgeons and covering a range of specimen complexity were taken from our hospital files. Questionnaires based on these cases were administered open-book-examination style to surgical attending physicians and trainees during surgical conferences at an academic medical center. MAIN OUTCOME MEASURES: Scores from questionnaires. RESULTS: Surgeons misunderstood pathologists' reports 30% of the time. Surgical experience reduced but did not eliminate the problem. Streamlined report formatting exacerbated the problem. CONCLUSIONS: A communication gap exists between pathologists and surgeons. Familiarity with report format and clinical experience help reduce this gap. Paradoxically, stylistic improvements to report formatting can interfere with comprehension and increase the number of misunderstandings. Further investigation is required to reduce the number of misunderstandings and, thus, medical errors.     

Identification of trypanosomes isolated from reduviidae from North Chile

Examination of 54Triatoma infestans from a village near the European Southern Observatory La Silla in Chile and of 9Triatoma spinolai from the territory of the observatory showed that 10T. infestans were infected with trypanosomatids. Mice were infected with in vitro cultures initiated with five different trypanosomatid isolates and treated with the immunosuppressive drug cyclophosphamide to increase the parasitemia of the flagellates. Evidence of the presence ofT. cruzi was provided by a comparative biometrical analysis of blood trypomastigotes and the occurrence of intracellular amastigotes. Three methods for further identification were used: examination of kDNA ultrastructure, disc electrophoresis of soluble proteins and theAaptos papillata II lectin induced agglutination. We obtained the following results for all isolates: (1) presence of a central band of the kDNA; (2)T. cruzi specific double bands of the protein patterns; (3) positive reaction withAaptos papillata II. No differences between the five isolates from Chile andT. cruzi orT. cruzi-like strains from other countries could be observed. Based on these results an infection of the bugs withT. rangeli andT. conorhini could be excluded.Supported by BMZ, Research Program FIOCRUZ, Rio de Janeiro/BNI, Hamburg     

“Far from the heart far from the eye”: Evidence from the Capgras delusion

Introduction. Capgras syndrome is characterised by the belief that a significant other has been replaced by an identical-looking impostor. These patients have no difficulties with visual recognition but fail to show a skin conductance response (SCR) to the objects of the delusion. A case of Capgras delusion (YY), specifically characterised by the absence of brain lesions, constituted a good opportunity to test the relationship between SCR hyporesponsiveness and eye movement patterns to familiar and unfamiliar faces.

Methods. Visual scan path and SCR were recorded for YY and 8 controls during the presentation of family members’ photographs matched with unfamiliar faces of the same sex, age, and physical likeness. Eye movement patterns were explored by selecting three specific areas of interest (AOI) involving the eyes, the mouth, and the face regions.

Results. In contrast with controls, YY showed a reduction in number and sum of fixation durations to the eyes (p<.01) and no differential SCRs (p>.05) to familiar vs. nonfamiliar faces. SCR and fixation duration to family members’ eyes were significantly correlated (r=.77) in both YY and controls.

Conclusions. Eye region exploration seems to be related to the autonomic reactivity elicited by the affective valence of familiar faces.